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Encompassing Amino Acids: VHL: 2-212(end); CUL2: 2-745(end); ELOB: 2-118; ELOC: 2-112(end); Rbx1: 2-108(end)
Application: Useful for the study of protein degradation and PROTAC design, and selectivity profiling.
Assay Conditions: ARV-771-mediated binding of VHL BPS #100373 to BET bromodomain was assessed using A-screen technology. Various amounts of VHL were incubated in 10-ul reaction with ARV-771 and GSTBRD3(BD2) BPS #31033 at room temperature for one hour. After this, FLAG-acceptor beads and GSH-donor beads were added followed by detection of A-counts.
Description: VHL/CUL2/ELOB/ELOC/RBX1 complex of human VHL (GenBank Accession No. NM_000551) a.a. 2-212(end) with N-terminal FLAG-tag and MW= 25 kDa, human CUL2 (GenBank Accession No. NM_003591), a.a. 2-745(end) with N-terminal FLAG-tag and MW= 88 kDa, human ELOB (GenBank Accession No. NM_007108), a.a. 2-118 with N-terminal FLAG-tag and MW= 14 kDa, ELOC (GenBank Accession No. NM_005648), a.a. 2-112(end) with N-terminal His-tag and MW= 13 kDa human Rbx1 (GenBank Accession No. NM_014248), a.a. 2-108(end) with N-terminal His-tag and MW= 13 kDa, individually expressed in a HEK293 expression system.
Format: Aqueous buffer solution
Formulation: 40 mM Tris-HCl, pH 8.0, 110 mM NaCl, 2.2 mM KCl, 20% glycerol, 3 mM DTT.
Genbank: VHL: NM_000551; CUL2: NM_003591; ELOB: NM_007108; ELOC: NM_005648; Rbx1: NM_005648
Purity: ≥80%
Storage Stability: At least 6 months at -80°C.
Supplied As: Aqueous buffer solution.
Tags: VHL: N-terminal FLAG-tag; CUL2: N-terminal FLAG-tag; ELOB: N-terminal FLAG-tag; ELOC: N-terminal His-tag; Rbx1: N-terminal His-tag
Uniprot: VHL: P40337; CUL2: Q13617; ELOB: Q15370; ELOC: Q15369; Rbx1: P62877
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Kamura, T., et al., Science. 1999; 284(5414): 657-661.
2. Stebbins, C.E., et al., Science. 1999; 284(5413): 455-461.
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